Preparation of pH-sensitive polymer by thermal initiation polymerization and its application in fluorescence immunoassay

A fluorescence immunoassay for human IgG (Ag) was developed using a pH-sensitive polymer prepared by thermal initiation or redox initiation polymerization as a carrier. In the competitive immunoassay, appropriate quantity of Ag was immobilized on the polymer and the standard Ag (or sample) solution, and a constant amount of fluorescein isothiocyanate labeled goat anti-human IgG antibody (Ab-FITC) was added. Immobilized Ag and the standard (or sample) Ag competed for binding to the Ab-FITC in 37 C in homogeneous format. After changing the pH to separate the polymer-immune complex precipitate, it was re-dissolved and determined by fluorescence method. The results showed that the immobilization efficiency, immunological reaction activities of immobilized Au and phase transition pH range were improved as Ag was immobilized by thermal initiation instead of redox initiation polymerization. Under optimum conditions, the calibration graphs for the Ag in both methods, thermal initiation and redox initiation, were linear over the concentration range of 0.0-1000 ng mL(-1), with detection limits 8 (thermal initiation) and 12 ng mL(1) (redox initiation), respectively. Moreover, some pH-sensitive polymer prepared only in organic solvent or under high temperature could also be used as an immunoreaction carrier by thermal initiation polymerization. Thermal initiation polymerization was a better immobilization mode. (C) 2004 Elsevier B.V. All rights reserved.

Identification and characterization of Bacillus anthracis by multiplex PCR on DNA chip

Bacillus anthracis can be identified by detecting virulence factor genes located on two plasmids, pXO1 and pXO2. Combining multiplex PCR with arrayed anchored primer PCR and biotin-avidin alkaline phosphatase indicator system, we developed a qualitative DNA chip method for characterization of B. anthracis, and simultaneous confirmation of the species identity independent of plasmid contents. The assay amplifies pag gene (in pXO1), cap gene (in pXO2) and Ba813 gene (a B. anthracis specific chromosomal marker), and the results were indicated by an easy-to-read profile based on the color reaction of alkaline phosphatase. About 1 pg of specific DNA fragments on the chip wells could be detected after PCR. With the proposed method, the avirulent (pXO1(+)/2(-), pXO1(-)/2(+) and pXO1(-)/2(-)) strains of B. anthracis and distinguished 'anthrax-like' strains from other B. cereus group bacteria were unambiguously identified, while the genera other than Bacillus gave no positive signal. (C) 2004 Elsevier B.V. All rights reserved.

Determination of phthalate acid esters plasticizers in plastic by ultrasonic solvent extraction combined with solid-phase microextraction using calix[4]arene fiber

Ultrasonic solvent extraction combined with solid-phase microextraction (SPME) with calix[4]arene/hydroxy-terminated silicone (C[4]/OHTSO) oil coated fiber was used to extract phthalate acid esters (PAEs) plasticizers in plastic, such as blood bags, transfusion tubing, food packaging bag, and mineral water bottle for analysis by gas chromatography (GC). Both extraction parameters (i.e. extraction time, extraction temperature, ionic strength) and conditions of the thermal desorption in a GC injector were optimized by analysis of eight phthalates. The fiber shows wonderful sensitivity and selectivity to the tested compounds. Owing to its high thermal stability (380 degreesC), the carryover effect that often encountered when using conventional fibers can be reduced by appropriately enhancing the injector temperature. The method showed linear response over two to four orders of magnitude with correlation coefficients (r) better than 0.996, and limits of detection (LOD) ranged between 0.006 and 0.084 mug l(-1). The relative standard deviation values obtained were less than or equal to 10%. bis-2-Ethylhexyl phthalate (DEHP) was the sole analyte detected in these plastics and recoveries were in the ranges 95.5-101.4% in all the samples. (C) 2004 Elsevier B.V. All rights reserved.

Oligonucleotide ligation assay-based DNA chip for multiplex detection of single nucleotide polymorphism

An oligonucleotide ligation assay-based DNA chip has been developed to detect single nucleotide polymorphism. Synthesized nonamers, complementary to the flanking sequences of the mutation sites in target DNA, were immobilized onto glass slides through disulfide bonds on their 5' terminus. Allele-specific pentamers annealed adjacent to the nonamers on the complementary target DNA, containing 5'-phosphate groups and biotin labeled 3'-ends, were mixed with the target DNA in tube. Ligation reactions between nonamers and pentamers were carried out on chips in the presence of T4 DNA ligase. Ligation products were directly visualized on chips through enzyme-linked assay. The effect of G:T mismatch at different positions of pentamers on the ligation were evaluated. The results showed that any mismatch between pentamer and the target DNA could lead to the decrease of ligation, which can be detected easily. The established approach was further used for multiplex detection of mutations in rpoB gene of rifampin-resistant Mycobacterium tuberculosis clinical isolates. (C) 2003 Elsevier B.V. All rights reserved.

Application and validation of a new biotic index using data from several water systems

Protozoans of Lake Donghu were collected from five stations using the PFU method. The sampling was conducted for one year and two times a month. The aim of this research was to test the applicability of a new protozoa biotic index, species pollution value (SPV) and community pollution value (CPV), established by the authors using data from the River Hanjiang. Each station's CPV was calculated from the SPV and the correlation analysis between the CPV and the comprehensive chemical index of stations I, II, III showed a significant correlation between them. The pollution status of the five stations was correctly evaluated by the CPV. These results suggested that the biotic index could be applied in water systems other than the River Hanjiang. The SPV of some protozoa species in Lake Donghu, not observed in the River Hanjiang were established. In order to further test the applicability of the biotic index, protozoan and chemistry data from the Rivers Torrente Stirone and Parma of Italy were used. The results showed that the CPV for the two rivers had a close relationship with the chemical water quality, which indicated that the biotic index could be applied in other parts of the world for the monitoring and estimating of water quality. Since the results of testing and verifying the biotic index in some other water systems in China were also satisfactory, this indicated that the biotic index has an extensive suitability for freshwater ecosystems. As long as more than 50% of the species in a sample have a SPV, the CPV calculated from the SPV is reliable for monitoring and evaluating water quality.

Determination of some reactive oxygen species scavengers based on the peroxidase-oxidase oscillator

This paper reports a new method for detection of ROS scavengers including superoxide dismutase, ascorbic acid and glutathione based on a 'probe' of peroxidase-oxidase biochemical oscillator. The oscillation period and amplitude change with different concentrations of scavengers. The linear ranges of superoxide dismutase, ascorbic acid and glutathione are respectively 1.56 x 10(-4)-1.56 x 10(-3) mg mL(-1), 1.75 x 10(-7) -1.75 x 10(-5) mol L-1 and 9.38 x 10(-7) -7.5 x 10(-5) mol L-1. The selectivity, linearity and precision for superoxide dismutase, ascorbic acid, and glutathione are presented and discussed. The results compared well with other standard methods for determination of superoxide dismutase, ascorbic acid and glutathione. Some possible steps in the overall reaction mechanisms are discussed.

Microcalorimetric studies on the thermogenesis of energy release of mitochondria isolated from rice

After isolation from rice mitochondria still have activity and can live for a long time by using its stored nutrients. The thermogenesis curves of energy release of the mitochondria isolated from variant strains of rice have been determined by using an LKB2277 bioactivity monitor. The differences in shape of the curves and the thermodynamic and kinetic characteristics of the thermogenesis of the mitochondria have been compared. The thermodynamic and kinetic parameters of energy release of the mitochondria in the thermogenesis increasing stage have been calculated, and the experimental thermokinetic equations of the thermogenesis have been established. (C) 2001 Elsevier Science B.V. All rights reserved.

Determination of trace level microcystins in water using solid-phase extraction and high performance liquid chromatography

This paper presents a method for trace level analysis of microcystins in water using solid-phase extraction and high performance liquid chromatography. The optimized condition enabled the determination of common microcystins at levels as low as 0.02 similar to 0.05 mug/L, and the liner range is from 0.1 mug/L to 50 mug/L. The method has been applied to the analysis of field sample from Dianchi lake.

Analysis of organchlorine compounds in water by solid phase microextraction and gas chromatography

In this work, both,solid phase microextraction (SPME) and solid phase extraction(SPE) were used to enrich organochlorine compounds in water samples and analyzed by gas chromatography with electron capture detector. The operating conditions of SPME have been studied and different kinds of solid phase were compared. Linear alkybenzene sulfonate(LAS) was added to the samples to investigate its effect on the analysis. The results indicated that polyacrylate was better than other commercial solid phases in extraction of moderated polar organic compounds and the sensitivity of SPME was higher than SPE. LAS affect much in liquid-liquid extraction and headspace SPME; but it has little effect on SPE and direct-SPME method. The applications showed that SPME was a fast and effective method in sample preparation.

Sol-gel method for the preparation of solid-phase microextraction fibers

A novel sol-gel method is applied for the preparation of solid-phase microextraction (SPME) fibers. Scanning electron microscopy experiments suggested a porous structure for the poly(dimethylsiloxane) (PDMS) coating. SPME-GC analysis provided evidence that the sol-gel fibers have some advantages, such as high thermal stability, efficient extraction rates, high velocities of mass transfer, and spacious range of application.

Formation and modeling of disinfection by-products in drinking water of six cities in China

Funding and support for this project was provided by NSFC (Grant No. 40771015), and Key International Science and Technology Cooperation Projects (Grant No. 22007DFC20180). The authors also gratefully acknowledge the support of Key Projects in the National Science & Technology Pillar Program in the Eleventh Five-year Plan Period (Grant No. 2006BAD01B06-02). The authors thank the CDCs of Daqing, Beijing, Tianjin, Zhengzhou, Changsha and Shenzhen cities for field and laboratory technical support.

Improving Ship Detection with Polarimetric SAR based on Convolution between Co-polarization Channels

The convolution between co-polarization amplitude only data is studied to improve ship detection performance. The different statistical behaviors of ships and surrounding ocean are characterized a by two-dimensional convolution function (2D-CF) between different polarization channels. The convolution value of the ocean decreases relative to initial data, while that of ships increases. Therefore the contrast of ships to ocean is increased. The opposite variation trend of ocean and ships can distinguish the high intensity ocean clutter from ships' signatures. The new criterion can generally avoid mistaken detection by a constant false alarm rate detector. Our new ship detector is compared with other polarimetric approaches, and the results confirm the robustness of the proposed method.

An electrochemical DNA sensor based on electrodepositing aluminum ion films on stearic acid-modified carbon paste electrode and its application for the detection of specific sequences related to bar gene and CP4 Epsps gene

An electrochemical DNA biosensor was fabricated by immobilizing DNA probe on aluminum ion films that were electrodeposited on the surface of the stearic acid-modified carbon paste electrode (CPE). DNA immobilization and hybridization were characterized with cyclic voltammetry (CV) by using methylene blue (MB) as indicator. MB has a couple of well-defined voltammetric redox peaks at the CPE. The currents of redox peaks of MB decreased after depositing aluminum ion films on the CPE (Al(III)/CPE) and increased dramatically after immobilizing DNA probe (ssDNA/Al(III)/CPE). Hybridization of DNA probe led to a marked decrease of the peak currents of MB, which can be used to detect the target single-stranded DNA. The conditions for the preparation of Al(III)/CPE, and DNA immobilization and hybridization were optimized. The specific sequences related to bar transgene in the transgenic corn and the PCR amplification of CP4 epsps gene from the sample of transgenic roundup ready soybean were detected by differential pulse voltammetry (DPV) with this new electrochemical DNA biosensor. The difference between the peak currents of MB at ssDNA/Al(III)/CPE and that at hybridization DNA modified electrode (dsDNA/Al(III)/CPE) was applied to determine the Specific sequence related to the target bar gene with the dynamic range comprised between 1.0 X 10(-7) mol/L to 1.0 x 10(-4) mol/L. A detection limit of 2.25 x.10(-8) mol/L. of oligonucleotides can be estimated.

Product monitoring and quantitation of oligosaccharides composition in agar hydrolysates by precolumn labeling HPLC

Since the discovery of multiple bioactivities for agarobiose oligomers, a quantitative method has been in great need to monitor the agarobiose oligomers. This report demonstrates that agarobiose oligomers can be separated with high resolution in HPLC after introducing a-naphthylamine into compounds. Agarobiose oligomers ranged from biose to decaose were isolated by Sephadex column. HPLC analysis indicated that each oliomer could be quantified with good linearity and a low detection limit of 0.1-4 mug/ml. The chromatographic profiles of agaro-oligosaccharides with different hydrolysis modes (hydrochloride, citric acid, solid acid, and hydroxyl radical degradation) showed that agarobiose could be obtained more than 57.8% using solid acid mediated hydrolysis, while hydrochloride acid could degrade agar into a series of agaro-oligosaccharides from biose to decaose. The yield of oligosaccharides was low if hydrolyzed by citric acid. The Fenton degradation can increase the speed of hydrolysis, but the product was complex. (C) 2004 Elsevier B.V. All rights reserved.